E3S Web of Conferences 151, 01025 (2020) https://doi.org/10.1051/e3sconf/202015101025 st 1 ICVAES 2019

Occurrence of gigantica and spp in Aceh

Muhammad Hambal1*, Rizka Ayuni1, Henni Vanda2, Amiruddin Amiruddin3, and Farida Athaillah1 1 Laboratory of Parasitology, Faculty of Veterinary Medicine, Universitas Syiah Kuala, Indonesia 2 Laboratory of Pharmacology, Faculty of Veterinary Medicine, Universitas Syiah Kuala, Indonesia 3 Laboratory of Clinic, Faculty of Veterinary Medicine, Universitas Syiah Kuala, Indonesia

Abstract. Fasciola gigantica and Paramphistomum spp. are trematode helminth causing severe economic losses in cattle farming in Aceh Province, Indonesia. This study was conducted to examine the correlation between the prevalence of F. gigantica and Paramphistomum spp with the body condition and sex of Aceh cattle. In total, 103 cattle (50 males and 53 females) from an abattoir in Banda Aceh were used. The body condition score was recorded and the number of fluke eggs in feces was examined coproscopically. The results showed that F. gigantica prevalence was 41% and 72% in females and males, respectively, whereas, the prevalence of Paramphistomum spp in females and males was 81% and 72%, respectively. The average number of fasciola eggs was 2.55 eggs/ g feces and 2.75 eggs/ g feces in females and males, respectively, The average number of Paramphistomum spp eggs was 127.6 eggs/g feces and 36.8 eggs/ g feces in males, and female respectively. Based on the Body Condition Score (BCS), the prevalence of both trematodes was higher in the skinny cattle (BSC 2 and 3). This study established that the infection of Fasciola in BCS 3 was higher than BCS 2 and 4. Male cattle were more susceptible to infection than females.

Keywords: epidemiology, sex influence, Body condition score, F. gigantica, Paramphistomum spp.

1 Introduction decreased to 6 % - 19% when the cattle fed on concentrate feed. In Indonesia, rice cultivation is part of the culture, Fasciola gigantica and Paramphistomum spp. are the and the paddy fields are important habitat for Fasciola common trematode helminth infesting cattle and other and Paramphistomum metacercariae, a source infection of worldwide [1-3]. These worms cause severe this trematode. A special case where trematode economic losses in cattle farming due to weight loss, prevalence rates reached 90% on cattle grazing freely in reduced production, low fertility rates, and in some paddy fields has been reported [15]. cases death [4-5]. Several previous studies reported high A lot of previous studies focused on the infestation of morbidity and mortality in young and adult cattle due to the F. gigantica and Paramphistomum spp. in ruminants these parasitic outbreaks [6-7], delayed growth, , [2, 18-19]. Nevertheless, no study has evaluated the tissue damage, especially in liver and bile ducts, and correlation between body condition score (BCS) and reduction in productivity [1, 8]. The economic loss in trematode infection in Aceh cattle. The BCS is an Indonesia due to the infestation of F. gigantica and important practical method of assessing the body Paramphistomum spp. is more than USD =$ 3.2 billion condition of the since it is a simple indicator of annually [9]. fat reserves in the body [20]. The information about the In general, cattle are infected by trematode when they correlation between BCS and parasitic status is swallow metacercariae attached to the grass and water also essential for livestock management decision making. [10-11]. Moreover, the density of metacercariae is Moreover, the influence of sex on the prevalence and egg influenced by the population size of snails Bulinus spp. number of the F. gigantica and Paramphistomum spp and Planorbis spp. as the intermediate hosts. The infestation in Aceh cattle has not been studied previously. prevalence of paramphistomiasis and relates to This information is essential in preventing the disease several factors, among them the grazing system [12-13], caused by the aforementioned parasites. For this reason, nutrition status, climate, ecosystem, and genetic aspects this study was conducted to examine the effect of sex and [14]. According to Putra et al. [13], pasture quality body condition score on the prevalence of the F. gigantica influenced the number of infected by trematode, and Paramphistomum spp. in Aceh cattle. F. gigantica, and Paramphistomum spp. In cattle fed with forage, the prevalence rate reached 30%, but the value

* Corresponding author: [email protected] © The Authors, published by EDP Sciences. This is an- open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). E3S Web of Conferences 151, 01025 (2020) https://doi.org/10.1051/e3sconf/202015101025 st 1 ICVAES 2019

2 Materials and Methods spp is bluish-gray. The eggs were counted and recorded using a binocular microscope at 1000X magnification. 2.1 Collection of parasites 2.4 Statistical analysis A total of 103 Aceh cattle, consisting of 50 males and 53 females from a slaughterhouse in Banda Aceh City were The data were subjected to the student T-test to analyze selected to be examined for fasciolosis and the effect of sex and body condition score on the worms' paramphistomosis prevalence rate, body condition score prevalence and egg number, while the Chi-squared test (BCS) and intensity of trematode infection. The feces was used to compare the spread between males and were collected from rectum coproscopically and put into females at a confidence limit of 5%. a transparent plastic bag. All samples were labeled and 0 stored in a cooling container at a temperature of 5 C 3 Results and Discussion before the examination. 3.1 Results 2.2 Measurement of body condition score (BCS) The study showed that 28 (27%) of the cattle were The BCS is a parameter used to analyze the body positively infected by F. gigantica and Paramphistomum condition of an unit. In this study, it was carried spp and indicated a mixed infection of both parasites in out through observation and palpation on the body fat Aceh cattle. Besides, 37 cattle had BCS 2, 47 had BCS 3 deposits under the skin around the base of the tail, spine, and 19 had BCS 4 (Table 1). Generally, there was a and hip. Assessment of the body condition was performed positive correlation between prevalence and BSC, where visually and by palpation of fat deposits in cattle body the pervasiveness of parasites increased with an increase parts such as at the hind and quarter of the back, the in BCS. For example, the prevalence of Paramphistomum processus spinosus, processus spinosus to processus spp. at BCS 2 was 16%, but increased to 55% and 94% at transversus, tuber coxae (hooks) between the coxae tuber BCS 3 and BCS 4 respectively. The study also detected a and the ischiadicus tuber, the right and left tuber coxae, mixed infection of F. gigantica and Paramphistomum and the base of the tail to the ischiadicus tuber [21]. The spp. in Aceh cattle. The higher mixed infestation was score ranges from 1-5, where score 1 = very thin, 2 = lean, found in cattle with BCS 3 followed by BCS 2 (Table 1). 3 = medium, 4 = fat and 5 = very fat. The results of the stool examination showed the eggs of F. gigantica were golden yellow with a size of 180 μm 2.3 Examination of worm eggs in length and 95 μm width. The average number of eggs in males was 2.76 eggs gram-1 feces and 2.56 eggs gram- The modification of sedimentation method was used in 1 feces in females. The eggs of Paramphistomum spp. this study (15). A total of three grams of feces were was transparent and showed clear embryonal cells. The weighed and put into the mortar and diluted with 20 ml of egg size was 113-175 µm in length, and 73-100 µm width, water. The solution of Tween-80 was added and then and equipped with an operculum. The number of homogenized by a magnetic stirrer for 5 min. The solution Paramphistomum eggs was 127.57 and 36.78 eggs gram- was filtered using mess 300, and the filtered material was 1 feces in males and females, respectively. The student T- sprayed with water, refilled with 100 ml of water and then test showed the sex of cattle gave a significant effect on left for three minutes before the removal of the prevalence and the number of eggs of the F. gigantica and supernatant. This procedure was repeated twice. The Paramphistomum spp. (P<0.05). remaining sediment was further dropped with 1% The study showed the prevalence of F. gigantica in Methylene blue to distinguish debris from trematode eggs male cattle was higher than in females (Table 2). The and transferred into a petri dish that was given a line for contrary finding was found in Paramphistomum spp. In orientation. The presence of Fasciola sp. eggs is which the higher prevalence was recorded in female cattle characterized by golden yellow, while Paramphistomum (Table 2).

Tabel 1. Body condition score and trematode prevalence of 103 cattle BCS Total F. gigantica Paramphistomum spp. F. gigantica + Prevalence Prevalence Paramphistomum spp.

5 0 0 (0%) 0 (0%) 0 (0%) 4 19 4 (21%) 18 (94%) 3 (16%)

3 47 17 (36%) 26 (55%) 15 (32%) 2 37 4 (11%) 6 (16%) 10 (27%) 1 0 0 (0%) 0 (0%) 0 (0%) .

2 E3S Web of Conferences 151, 01025 (2020) https://doi.org/10.1051/e3sconf/202015101025 st 1 ICVAES 2019

3.2 Discussion age, sex, and breed, whilst intrinsic aspects include climate, environment, and farm management [22-23]. The study showed 25 and 50 cattle samples were infested Animal age is one of the most important factors in by F. gigantica and Paramphistomum respectively, while fasciolosis, where the cattle older than 12 months are the remaining 28 were infected by both parasites. The more susceptible to F. gigantica compared to younger infestation of helminth in cattle is influenced by both ones [24]. internal and external factors. The external factors include

Table 2. The number of eggs in feces and the prevalence of Fasciola gigantica in cattle Sex Sample Positive Total of Average eggs per Prevalence eggs gram feces (%) a a Male 50 36 99 2.75 72 a b Female 53 22 56 2.55 41 Total 103 58 155 2.65 56.3 a,b Different superscript in the same column showed a significant difference between male and female (P< 0.05)

Tabel 3. The average of eggs in feces and the prevalence of Paramphistomum spp. in cattle Sex Sample Positive Total eggs EpG Prevalence (%) Male 50 36 4593 127.6 a 72 a Female 53 43 1582 36.8 b 81 b Total 103 79 2038 25.82 76.7 a,bDifferent superscript in in the same column showed a significant difference between male and female (P< 0.05) EpG = egg per gram feces.

The prevalence of F. gigantica in this study was The number of parasite eggs in feces showed the higher than in other reports in Indonesia and other tropical male cattle (2.75 eggs/g of feces) had a slightly higher countries such as Borneo [25], West Java [26], Vietnam number than females (2.55 eggs g-1 of feces). The number [27], and Iran [28]. of eggs of F. gigantica was higher than in a previous Based on sex, the prevalence rate of F. gigantica in report by [26], who recorded 1.31 eggs/g feces in cattle in male cattle was significantly higher than in females. Borneo. According to [27], there were 49 eggs/g feces Therefore, it was assumed the female had a lower risk (11.2%) in West Java. This is because Aceh Province is factor to F. gigantica infection. This finding is in known to have a higher risk of F. gigantica infestation agreement with [12], who examined the intensity of F. region in Indonesia [15, 31]. gigantica in cattle and established that the male's infection The study revealed that the BCS 3 dominated in all rate was higher than in females. This condition was of the examined cattle. This indicated most of the cattle attributed to the hormonal system where the estrogen slaughtered in Banda Aceh municipal were quite lean. triggers properties of Reticulo Endothelial System (RES) This finding was in agreement with [8], who reported that in cells mediated through direct chemical interaction with the level of intensity and prevalence of Paramphistomum lymphoid and nonlymphoid tissue, resulting in the release spp. in of Ethiopia was higher in animals with of soluble immunoregulatory factors. These factors are poor body conditions. The study also showed the cattle produced by thymic epithelium in response to an estrogen sample with BCS 3 was the most infected by F. gigantica stimulus. Furthermore, according to [25], the phagocytic and Paramphistomum. This is, probably, related to age activity of the RES in animals may be measured by the stage, an assertion supported by [15], where cattle with number of particles present in the blood. However, it is BCS 3 were mainly 2-4 years old, while 1.5 to 2.5 years well-known that estrogen stimulates the level of RES in old mostly had BCS 2. The young cattle are generally the animal body. This hormone induces the blood free-roaming in a dry field and therefore most exposure to clearance rate and increases the number of phagocytes metacercariae. It was assumed that most of the young cells in the liver. By so doing, it enhances the immune cattle do not have adequate antibody [32]. On the basis of system of the female cattle. Besides, the females are rarely body condition score, female cattle were more susceptible used in plowing in the paddy field, especially in gravid to F. gigantica infestation than the males. Besides, the condition, and therefore they have less contact with weight loss in females was significantly higher than in metacercariae, a source of infection. On the contrary, [30] males. This is in agreement with [33] who reported that reported a higher prevalence in females than in males in female cattle in India were more susceptible to F. Bangladesh. This difference was attributed to the gigantica than the males. However, according to [34], F. physiological conditions of female cattle during gravid gigantica leads to weight loss in goat (Capra hircus), and lactation. It may also be due to the lack of dietary though it did not specify the difference between males and supplements for production which leads to a decrease in females in response to the parasite. the immunity of females. It is assumed that in malnutrition condition, estrogen interaction with RES is deactivated

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